ISSR分子标记对杧果实生苗父本的早期鉴定

[目的]采用ISSR分子标记技术对杧果实生苗进行亲本早期鉴定,为提高杧果育种效率提供参考.[方法]采用ISSR分子标记技术,筛选不同引物鉴定97-1实生苗的父本.[结果]采用优化的ISSR反应体系,从18条引物中筛选获得4条特异引物UBC-848、UBC-857、UBC-864和UBC-868,其扩增条带多、稳定性强、多态性高、背景清晰.利用引物UBC-848、UBC-857、UBC-864、UBC-868分别对四川红柁、紫花柁和97-1实生苗进行扩增,获得的扩增片段长度在100～2000bp,四川红杧与97-1实生苗、紫花杧与97-1实生苗的共有特异性条带约为180和380 bp、280和380 bp、180和680 bp、680和770bp.[结论]采用ISSR分子标记技术和引物UBC-848、UBC-857、UBC-864、UBC-868构建了杧果实生苗亲本鉴定体系,可鉴定出97-1实生苗的父本为四川红杧,97-1实生苗是紫花杧和四川红杧的杂交种.     

陕西关中某羊场羊口疮病毒的分离鉴定与基因序列分析

羊口疮疫病的暴发可严重影响畜牧的经济效益,为探明陕西地区该病的发病原因。本研究从陕西关中地区某羊场出现特征症状病羊嘴部痂皮中分离病原,经过感染MDBK细胞系,获得1株病毒,经形态学、理化学、PCR检测及病理组织学等方法鉴定,该病毒分离株为羊传染性脓疱病毒(Orf virus,ORFV),命名为ORFV-SXGZ。依据GenBank中发表的ORFV特异性B2L、F1L及VIR基因的核酸序列,设计并合成3对引物,应用PCR方法成功对ORFV-SXGZ毒株的B2L、F1L及VIR基因进行克隆,并将其基因序列及推导的氨基酸序列与其他不同来源ORFV分离株进行比较分析。序列分析结果表明,ORFV-SXGZ毒株的B2L、F1L和VIR基因与其他已发表ORFV毒株之间的核苷酸同源性分别为96.7%~99.6%、95.8%~97.9%和95.0%~99.3%,氨基酸同源性分别为95.3%~99.7%、95.8%~98.2%和95.5%~98.4%。系统进化树分析结果显示,ORFV-SXGZ毒株与国内新疆和甘肃等西北部地区分离株的亲缘关系更近,证实陕西关中地区养殖场中存在羊口疮病毒感染。     

A型口蹄疫病毒VP1蛋白间接ELISA检测方法的建立

以原核表达经纯化复性制备的A型口蹄疫病毒(FMDV)VP1蛋白代替传统病毒抗原包被,建立快速、安全、有效的A型FMDV抗体ELISA检测方法。对前期制备的A型VP1蛋白进行Western-Blot检测,结果表明,VP1蛋白能够特异性识别A型FMDV阳性牛血清,可作为检测抗原建立检测A型FMDV抗体的方法。以最佳质量浓度1mg/L VP1蛋白为检测抗原,方阵滴定法确定最佳检测血清稀释度为1∶50[V(血清)∶V(封闭液)],最佳的酶标二抗稀释度为1∶2 000[V(酶标二抗)∶V(封闭液)],建立A型FMDV抗体ELISA检测方法。该方法的敏感性为94.32%,特异性为99.09%,批内与批间重复试验变异系数均小于8%。采用该方法检测201份临床血清,与液相阻断ELISA试剂盒的符合率达92.54%。研制的VP1-ELISA试剂盒特异、敏感、稳定、操作简便,可用来监控A型口蹄疫抗体水平。     

部分肉牛类群CAPN1基因第9外显子多态性及其与肉质性状的相关性

通过研究部分肉牛类群CAPN1基因第9外显子多态性及其与肉质性状的相关性,发现与肉牛生产性状相关的分子标记,为加快肉牛选育进程,提高肉牛生产性能提供参考。采用PCR-SSCP技术对早胜牛(庆阳类群、平凉类群)、南杂牛(南德温牛×早胜牛庆阳类群)、秦杂牛(秦川牛×早胜牛平凉类群)、西杂牛(西门塔尔牛×早胜牛平凉类群)、荷斯坦奶公牛和澳洲黑牛共391头个体的CAPN1基因外显子9进行遗传变异检测,运用SPSS17.0软件分析早胜牛平凉类群遗传变异与肉质性状的关联性。结果表明,CAPN1基因第9外显子存在突变位点G458C,并检测到GG、GC和CC 3种基因型。相关性分析表明,CAPN1的G458C位点与pH、剪切力和滴水损失差异显著相关(P0.05),CC基因型剪切力和滴水损失显著低于GG和GC型(P0.05),其他性状在各基因型间差异不显著。因此,推测CAPN1基因的突变位点G458C可以作为牛肉嫩度的遗传标记。     

威宁黄牛SOCS1基因外显子区SNP与体尺性状关联性研究

为研究SOCS1基因突变对威宁黄牛生长性能的影响,以106头威宁黄牛为研究对象,采用SSCP、DNA测序方法分析SOCS1基因外显子区SNP与不同个体体尺性状的相关性。结果显示：A＋815C位点在威宁黄牛中有不同分型,在威宁黄牛公畜中,A＋815C位点与体高、体直长有极显著关联,CC基因型个体体高、体直长均值最大,为有利基因型,对胸深的影响达到显著性水平,具有杂合子优势;在母畜中,A＋815C位点对胸围的影响达到显著水平,杂合子 AC基因型个体胸围均值最大。     

九丰1号金银花定植早期最佳种植密度的研究

以九丰1号金银花为供试材料,设计了4种种植密度（株距：1.4、1.2、1.0、0.8 m,行距均为1.5 m）,密度范围为4 762-8 333株/hm2,探讨在淮安地区金银花种植早期的适宜种植密度。结果表明,在江苏淮安地区,九丰1号栽培的前3 a在试验设计的种植密度范围内,产量随种植密度的增加而增加;种植密度为8 333株/hm2的处理各年份的产量均高于其它密度处理,其中2011年、2012年和2010-2012年3 a合计产量与其它密度处理差异显著。     

维生素B1对中国对虾生长影响的研究

研究了中国对虾饵料中维生素Ｂ１的适宜含量，当１００ｇ饵料含６ｍｇＶｂ１时，中国对虾增长率和存活率最高；ＶＢ１缺乏或过量，都影响对虾的生长和存活，ＶＢ１在对虾组织中的含量随饵料中ＶＢ１含量的增加而增加，以６ｍｇＶＢ１／１００ｇ饵料最高，如过量欺一反而降。ＶＢ１可影响对虾肝胰脏α－淀粉酶的活性，当１００ｇ饵料含６ｍｇＶＢ１时，这种酶活性最高，对虾生长也最好。     

Influence of high dietary α-tocopherol intakes on specific immune response, nonspecific resistance factors and disease resistance of healthy and aflatoxin B1-induced immunocompromised Indian major carp, Labeo rohita (Hamilton)

The effect of aflatoxin treatment and/or feeding of a high level of α‐tocopherol on immune response and disease resistance was investigated in Indian major carp, Labeo rohita. Group A served as a healthy control, group B was treated with aflatoxin, group C was fed a high level of α‐tocopherol whereas group D was exposed both to aflatoxin and a high level of dietary α‐tocopherol for 60 days. Aflatoxin B₁ (AFB₁) was injected once intraperitoneally into fish on the first day of the experiment (groups B & D). High levels of DL‐α‐tocopherol (1000 mg kg^–1 feed) were provided to healthy as well as AFB₁‐treated immunocompromised fish for 60 days (groups C & D). At the end of the experiment blood samples were assayed for changes in nonspecific immunity and humoral protein levels. Disease resistance against two common bacterial pathogens viz., Aeromonas hydrophila and Edwardsiella tarda were evaluated in all groups. Significant (P < 0.05) suppression of specific immunity as measured through haemagglutination (HA) titre against sheep red blood cells (SRBCs) as well as bacterial (formalin‐killed E. tarda) agglutination titre; nonspecific resistance factors viz., globulin level, serum bactericidal and lysozyme activities, neutrophil activities, and disease resistance against two bacterial pathogens only in aflatoxin‐treated fish with respect to the control group, clearly indicated the immunosuppressive nature of aflatoxin. Feeding of a high level of α‐tocopherol to AFB₁‐treated immunocompromised fish significantly (P < 0.05) raised specific immunity, nonspecific resistance factors and disease resistance capacity when compared with aflatoxin‐exposed fish. Disease resistance and enhancement of immune status through feeding of high levels of α‐tocopherol to healthy as well as AFB₁‐treated immunocompromised fish confirmed the potential of α‐tocopherol in carp feed for prevention of disease and for combating natural/environmental immunosuppressants.     

Neuroimmunological regulation of α-MSH release in tilapia (Oreochromis mossambicus)

This study describes the effects of IL-1 (interleukin 1) and LPS (bacterial endotoxin lipopolysaccharide) on the release of α-MSH (alpha melanocyte stimulating hormone) from the neurointermediate lobe (NIL) of the teleost Oreochromis mossambicus (tilapia). In vivo treatment of tilapia with IL-1 for 8 days led to a 49% inhibition of basal α-MSH release, measured by means of an in vitro micro-superfusion technique. The treatment did not affect the sensitivity of the tissue to TRH. In vitro, the release of α-MSH was inhibited by LPS in a dose dependent manner. In addition to its effects on the unstimulated release of the hormone, LPS also blunted the response to a TRH stimulation. Together with recent results obtained by others demonstrating the effects of (neuro-)peptides on immune parameters and the presence of cytokines in fish, the present data establish the bidirectional character of the communication between the immune and the (neuro-)endocrine systems in teleosts.

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Résumé Cette étude décrit les effets de l'IL-1 (interleukin 1) et du LPS (bacterial endotoxin lipopolysaccharide) sur la libération de α-MSH (alpha melanocyte stimulating hormone) par le lobe neurointermédiaire (NIL) d'un téléosteen Oreochromis mossambicus (tilapia). Le traitement in vivo du tilapia avec l'IL-1 pendent 8 jours conduit à une inhibition de 49% de la libération basale d'α-MSH mesurée à l'aide d'une technique in vitro de micro-superfusion. Ce traitement ne modifie pas la sensibilité du tissue au TRH. In vitro, la libération de α-MSH est inhibée par le LPS de manière dose-dépendante. En plus de ses effets sur la libération basale de α-MSH, le LPS bloque aussi la réponse à une stimulation par le TRH. Confrontés à des donnés récemment publiés montrant les effets de (neuro-)peptides sur les paramêtres immunitaires et la présence de cytokines chez les poissons, nos résultats établissent le caractère bidirectionnel de la communication entre le système immunitaire et les systèmes (neuro-)endocriniens.

     

The effect of stocking size on the first winter survival of whitefish, Coregonus lavaretus (L.), in the Gulf of Bothnia, Baltic Sea

The anadromous whitefish, Coregonus lavaretus (L.), is the most numerous fish species stocked in the Gulf of Bothnia, Baltic Sea. One-summer-old-whitefish fingerlings are mostly 8–10 cm long when released annually in September–October, whereas the wild whitefish are 10–12 cm at that time. About 6 million, one-summer-old, spray-marked, whitefish were released in the northern and central parts of the Gulf in 1995–1998. To study the effect of the stocking length on the survival of the marked fish, the length of the recaptured whitefish as 1-year-olds was back-calculated. Altogether 1106 whitefish recaptured in the Gulf of Bothnia were analysed. The back-calculated length was slightly greater than the stocking length but not as large as the length of the wild fish. In the central part of the Gulf of Bothnia, where the mean stocking length was more than 10 cm, the back-calculated length was 10.5–11.1 cm. In the northern part of the Gulf the mean stocking length varied between 8.8 and 10.0 cm annually, and the corresponding back-calculated mean lengths were 9.3–9.7 cm. It also seemed that bigger fingerlings started their feeding migration earlier or they migrated faster than the smaller ones to the southern parts of the Gulf of Bothnia.